Exo70E2 is essential for exocyst subunit recruitment and EXPO formation in both plants and animals

被引:71
作者
Ding, Yu [1 ,2 ]
Wang, Juan [1 ,2 ]
Lai, John Ho Chun [1 ,2 ,3 ,4 ]
Chan, Vivian Hoi Ling [1 ,2 ]
Wang, Xiangfeng [1 ,2 ]
Cai, Yi [1 ]
Tan, Xiaoyun [5 ]
Bao, Yiqun [5 ]
Xia, Jun [3 ,4 ]
Robinson, David G. [6 ]
Jiang, Liwen [1 ,2 ,7 ]
机构
[1] Chinese Univ Hong Kong, Sch Life Sci, Ctr Cell & Dev Biol, Shatin, Hong Kong, Peoples R China
[2] Chinese Univ Hong Kong, State Key Lab Agrobiotechnol, Shatin, Hong Kong, Peoples R China
[3] Hong Kong Univ Sci & Technol, Div Life Sci, Div Biomed Engn, Kowloon, Hong Kong, Peoples R China
[4] Hong Kong Univ Sci & Technol, State Key Lab Mol Neurosci, Kowloon, Hong Kong, Peoples R China
[5] Nanjing Agr Univ, Coll Life Sci, Nanjing 210095, Jiangsu, Peoples R China
[6] Heidelberg Univ, Ctr Organismal Studies, Dept Plant Cell Biol, D-69120 Heidelberg, Germany
[7] Chinese Univ Hong Kong, Shenzhen Res Inst, Shenzhen 518057, Peoples R China
基金
中国国家自然科学基金;
关键词
SUSPENSION-CULTURED CELLS; ENDOPLASMIC-RETICULUM EXPORT; VACUOLAR SORTING RECEPTORS; POLLEN-TUBE GROWTH; TOBACCO BY-2 CELLS; PLASMA-MEMBRANE; ARABIDOPSIS-THALIANA; SUBCELLULAR-LOCALIZATION; MULTIVESICULAR BODIES; TETHERING FACTORS;
D O I
10.1091/mbc.E13-10-0586
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
In contrast to a single copy of Exo70 in yeast and mammals, the Arabidopsis genome contains 23 paralogues of Exo70 (AtExo70). Using AtExo70E2 and its GFP fusion as probes, we recently identified a novel double-membrane organelle termed exocyst-positive organelle (EXPO) that mediates an unconventional protein secretion in plant cells. Here we further demonstrate that AtExo70E2 is essential for exocyst subunit recruitment and for EXPO formation in both plants and animals. By performing transient expression in Arabidopsis protoplasts, we established that a number of exocyst subunits (especially the members of the Sec family) are unable to be recruited to EXPO in the absence of AtExo70E2. The paralogue AtExo70A1 is unable to substitute for AtExo70E2 in this regard. Fluorescence resonance energy transfer assay and bimolecular fluorescence complementation analyses confirm the interaction between AtExo70E2 and Sec6 and Sec10. AtExo70E2, but not its yeast counterpart, is also capable of inducing EXPO formation in an animal cell line (HEK293A cells). Electron microscopy confirms the presence of double-membraned, EXPO-like structures in HEK293A cells expressing AtExo70E2. Inversely, neither human nor yeast Exo70 homologues cause the formation of EXPO in Arabidopsis protoplasts. These results point to a specific and crucial role for AtExo70E2 in EXPO formation.
引用
收藏
页码:412 / 426
页数:15
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